RESUMEN
Brain function in normal aging and neurological diseases has long been a subject of interest. With current technology, it is possible to go beyond descriptive analyses to characterize brain cell populations at the molecular level. However, the brain comprises over 100 billion highly specialized cells, and it is a challenge to discriminate different cell groups for analyses. Isolating intact neurons is not feasible with traditional methods, such as tissue homogenization techniques. The advent of laser microdissection techniques promises to overcome previous limitations in the isolation of specific cells. Here, we provide a detailed protocol for isolating and analyzing neurons from postmortem human brain tissue samples. We describe a workflow for successfully freezing, sectioning and staining tissue for laser microdissection. This protocol was validated by mass spectrometric analysis. Isolated neurons can also be employed for western blotting or PCR. This protocol will enable further examinations of brain cell-specific molecular pathways and aid in elucidating distinct brain functions.
Asunto(s)
Encéfalo/citología , Neuronas/metabolismo , Proteoma/metabolismo , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Captura por Microdisección con Láser , Masculino , Persona de Mediana Edad , Cambios Post Mortem , Espectrometría de Masas en TándemRESUMEN
The HUPO Brain Proteome Project (HUPO BPP) held its 17(th) workshop in Sao Paulo, Brazil, on May 24 and 25, 2012. The focus was on the progress on the Human Brain Proteome Atlas as well as ideas, strategies and methodological aspects in clinical neuroproteomics.
Asunto(s)
Biomarcadores/análisis , Química Encefálica , Proteínas del Tejido Nervioso/análisis , Proteoma/análisis , Proteómica/métodos , Animales , Investigación Biomédica , HumanosRESUMEN
The Human Brain Proteome Project (HUPO BPP) aims at advancing knowledge and the understanding of neurodiseases and aging with the purpose of identifying prognostic and diagnostic biomarkers, as well as to push new diagnostic approaches and medications. The participating groups meet in semi-annual workshops to discuss the progress, as well as the needs, within the field of proteomics. The 9(th) HUPO BPP workshop took place in Barbados from 9-10 January, 2008. Discussing the future HUPO BPP Roadmap, the attendees drafted the so called HUPO BPP wish list containing timelines, suggestions and missions. This wish list will be updated regularly and will serve as a guideline for the next phase.